Modular radiochemistry synthesis system

ABSTRACT

A modular chemical production system includes multiple modules for performing a chemical reaction, particularly of radiochemical compounds, from a remote location. One embodiment comprises a reaction vessel including a moveable heat source with the position thereof relative to the reaction vessel being controllable from a remote position. Alternatively the heat source may be fixed in location and the reaction vial is moveable into and out of the heat source. The reaction vessel has one or more sealing plugs, the positioning of which in relationship to the reaction vessel is controllable from a remote position. Also the one or more reaction vessel sealing plugs can include one or more conduits there through for delivery of reactants, gases at atmospheric or an elevated pressure, inert gases, drawing a vacuum and removal of reaction end products to and from the reaction vial, the reaction vial with sealing plug in position being operable at elevated pressures. The modular chemical production system is assembled from modules which can each include operating condition sensors and controllers configured for monitoring and controlling the individual modules and the assembled system from a remote position. Other modules include, but are not limited to a Reagent Storage and Delivery Module, a Cartridge Purification Module, a Microwave Reaction Module, an External QC/Analysis/Purification Interface Module, an Aliquotting Module, an F-18 Drying Module, a Concentration Module, a Radiation Counting Module, and a Capillary Reactor Module.

RELATED APPLICATION

This Application is a continuation of U.S. patent application Ser. No.14/582,885 filed on Dec. 24, 2014, now U.S. Pat. No. 9,211,520, whichitself is a divisional of and claims priority to U.S. patent applicationSer. No. 13/058,526 filed on Jun. 2, 2011 now issued as U.S. Pat. No.8,951,480, which itself claims priority to PCT Application NoPCT/US2009/004745 filed Aug. 19, 2009 which claims priority to U.S.Application No. 61/090,152 filed on Aug. 19, 2008. The contents of theaforementioned applications are hereby incorporated herein by referencein their entirety. Priority to the aforementioned applications arehereby expressly claimed in accordance with 35 U.S.C. §§119, 120, 365and 371 and any other applicable statutes.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

This invention was made with Government support under Grant No.DE-FG02-06ER64249, awarded by the U.S. Department of Energy, and GrantNo. CA086306, awarded by the National Institute of Health. TheGovernment has certain rights in the invention.

FIELD OF INVENTION

Modular, automated chemical synthesis apparatus suitable for thepreparation of small quantities of chemicals, particularly radiopharmaceuticals are described. The system and apparatus are also usablefor preparing other compounds which may unstable or are desired in smallquantities produced at the location they will be used.

BACKGROUND

Radiochemistry is a complex area of chemistry that is an increasinglyimportant part of providing diagnostic imaging in the clinical setting.The growth in Positron Emission Tomography (PET) and Single-PhotonEmission Computed Tomography (SPECT) means that researchers and clinicalscientists are highly interested in synthesizing new diagnosticcompounds and perfecting synthesis techniques for new radioisotopes,“tracers”, “probes” and “biomarkers”. However, because of theradioactive decay of the prepared materials, the hazard of radiationexposure to medical personnel, and the chemical instability of theradiolabeled materials, these radiation labelled compounds mustgenerally be prepared on site and the diagnostic procedure conductedwithin a short period of time after the materials are prepared.

Radiochemistry has traditionally required manually intensive, bench topmanipulation of chemicals with fairly standard chemical apparatus withinan environment that is designed to protect the chemist from exposure ofthe fingers, hands, or body to radiation. Low-dose radiochemistry (<acouple of mCi) can be conducted in an appropriately reinforced fume hoodwith lead bricks and other types of passive shielding. High dose(curies) synthesis must be conducted in a hot cell with considerablyhigher shielding and safety requirements.

Manually-operated assemblies of reaction vessels, sensors, heaters, etc.are commonplace. Automated radiochemistry devices also exist (e.g.,commercial FDG, methylation, etc.). However these devices areessentially optimized for a specific chemistry process and are not userconfigurable without having to physically manipulate hardware andreprogram the synthesis process. Existing radiochemical reaction systemsare also generally not capable of performing high-pressure reactions(e.g. >50 psi).

A typical prior manual radiochemistry setup for performingradiochemistry experiments and synthesis of diagnostic materialscomprises digitally-controlled hotplates and oil baths within a hot cellmade of lead-bricks. Syntheses are generally followed by standard manualpurification procedures.

Whether a low dose or high dose environment is involved, the increasinguse of radiochemistry to perform synthesis with a variety of isotopes,including but not limited to ¹⁸F, ¹¹C, ¹³N, ¹⁵O, ¹²³I, ¹²⁴I, ⁶⁴Cu, ⁶⁸Ga,etc. means that there is an increasing risk of radiation exposure to thechemist. Automated radiochemistry units are available, and severaldevices referred to as “automated synthesis modules” exist for specifictypes of reactions that are routinely and repeatedly conducted,including electrophilic chemistry, nucleophilic chemistry andmethylation. However, these units are typically hard wired with a fixedcomponent configuration for a specific number of reaction steps, solutevolumes, and radiation levels and are notably inflexible for theexperimental chemist or to handle multiple different end products. Suchunits typically are “black boxes” with pushbutton operation and must bephysically rewired and the hardware and software reconfigured to performa new or different synthesis step. This is in marked contrast to thevisual and interactive prior art bench top manual apparatus.

SUMMARY

The modular radiosynthesis system which incorporates features of theinvention consists of a sequence of subsystems or “modules”, each ofwhich performs a unit operation.

Each module includes control and telemetry capability so that it can beremotely controlled and monitored in a stand-alone fashion as well asreadily assembled into a system to perform different reaction protocols.Stand-alone operation permits straightforward “plug-and-play”reconfiguration of modules without reprogramming, and requiring onlyfluid connections be made between modules to implement the desiredsequences.

Modules are preferrably constructed with a deep, thin profile so theycan be stacked side by side in a compact space such as a mini-cell,while retaining an intuitive relationship between physical positions ofmodules and the sequence of steps in the radiochemical synthesis. Otherstacking arrangements are also possible.

The apparatus enables extremely flexible chemistry to be conducted,“recorded” electronically and the replayed back in an automated fashion,including driving multiple units and some unique telemetry functions.The apparatus has particular utility as a research radiochemistryplatform to provide easy synthesis of a wide range of compounds withlittle or no radiation exposure to the equipment operator. Because theplatform is entirely remotely controllable, it can easily be configuredto controllers and/or software for either manual or automated operation.Thus, the same platform can be used for synthesis development usingmanual control as well as for routine production by automatic control,thereby saving considerable development time that is normally needed totranslate the optimized manual synthesis from a manual apparatus to anautomated system.

While the description below is directed to a system incorporatingvarious unit-operation modules to perform a complete synthesis of apurified end product ready for clinical use, including modules forstoring and feeding in a controlled manner all of the required reactantsand reagents, the modules themselves are unique devices, several ofwhich are independently patentable.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a schematic representation of a first embodiment of a roboticsynthesis module also referred to as a reaction module, in its startingconfiguration incorporating features of the invention.

FIG. 2 is a schematic representation showing the configuration of therobotic synthesis module of FIG. 1 during a representative reactionunder pressure.

FIG. 3 is a schematic representation showing the configuration of therobotic synthesis module of FIG. 1 during a representative reactionsolvent evaporation stage.

FIG. 4 is a schematic representation showing the configuration of therobotic synthesis module of FIG. 1 during a product transfer, which mayalso include flushing and washing out of the reaction vessel.

FIG. 5 is a schematic diagram of the single reactor unit of FIGS. 1-4with the reaction vessel connected to various operational components andmaterial feed sources.

FIG. 6 is a schematic diagram of a single Solid Phase Extraction (SPE)module.

FIG. 7 is a schematic diagram illustrating operation in series of threeSPE module.

FIG. 8 is a schematic diagram illustrating modularity with multipleindividual reaction units arranged serially to provide an N-stepreaction module.

FIG. 9 is a schematic representation of one embodiment of a reagentstorage and delivery module.

FIGS. 10 and 11 are schematic of a representations of two embodiments ofliquid transfer subsystems.

FIG. 12 is a schematic representation of one embodiment of a cartridgepurification module.

FIG. 13 is a schematic representation of one embodiment of a highpressure robotic reaction module.

FIGS. 14, 15, 16 and 17 are schematic representations showing a moveablehigh pressure robotic reaction module in 4 alternative locations.

FIG. 18 is a schematic representation of a microwave reaction module.

FIG. 19 is a schematic representation of an analytical QC orpurification system interface module.

FIG. 20 is a schematic representation of an aliquotting module.

FIG. 21 is a schematic representation of a concentration module.

FIGS. 22, 23 and 24 are schematic representations of embodiments of aradiation counting module.

FIG. 25 is a schematic representation of a capillary or microfluidicreaction module.

FIG. 26 is a schematic representation of a multiple module reactionsystem.

FIG. 27 is a schematic representation of a first reaction module in thefluorination step in a manual synthesis of [¹⁸F] FAC.

FIG. 28 is a schematic representation of a second reaction module in thebromination and cytosine coupling step in a manual synthesis of [¹⁸F]FAC.

FIG. 29 is a schematic representation of a third reaction module in thedeprotection step in a manual synthesis of [¹⁸F] FAC.

FIG. 30 is a schematic representation of the fluorination step, using afirst modular system incorporating features of the invention, in therobotic synthesis of [¹⁸F] FAC.

FIG. 31 is a schematic representation of the bromination and cytosinecoupling step using a second modular system incorporating features ofthe invention, in the robotic synthesis of [¹⁸F] FAC.

FIG. 32 is a schematic representation of the deprotection step using athird modular system incorporating features of the invention, in therobotic synthesis of [¹⁸F] FAC.

FIG. 33 is a radio-HPLC spectrum of [¹⁸F] FNB produced using a modularsystem incorporating features of the invention.

FIGS. 34, 35, and 36 are radio-TLC spectrums for three stages in theproduction of [¹⁸F] FDG with FIG. 34 showing the spectrum afterfluorination, FIG. 35 showing the spectrum after hydrolysis and FIG. 36showing the spectrum after purification.

FIG. 37 is an HPLC chromatogram indicating the purity of D-[¹⁸F] FAC.

FIG. 38 is an HPLC chromatogram indicating the purity of L-[¹⁸F] FAC.

FIG. 39 is an HPLC chromatogram indicating the purity of L-[¹⁸F] FMAC.

FIG. 40 is an HPLC chromatogram indicating the purity of [¹⁸F]-CA.

DETAILED DESCRIPTION

Disclosed are modular component assemblies referred to herein as“plug-and-play modules” that allow an operator to replace the manual“touch and see” mode of experimental synthetic radiochemistry andradiolabeling with a unit that can be controlled remotely from aradiation protected environment in an automated and repeatable manner.Specific advantages of the modular system include, but are not limitedto:

-   -   a) A modular, n-stage robotics device that can be scaled        infinitely through the addition of “generic” reaction modules to        a provide a broad range of synthesis capability and address a        broad range of reaction complexity. Each module or component of        the system has its own control system and telemetry capability.    -   b) Incorporation of inexpensive but streamlined Solid-Phase        Extraction (SPE) units for coordinating purification and QC of        multi-step reactions.    -   c) Implementation of a unique robotic format that is both        relevant and familiar to the chemist, but also facilitates a        vessel pressurization function that supports high-pressure (>150        psi) reactions. No existing commercial automated synthesis units        are capable of performing high-pressure radiochemistry.    -   d) Utilization of a “remote control” unit that is not limited to        operation within a fume hood or hot cell. This unit allows        streamlined control of each robotic actuator and SPE component        utilizing, for example, cable telemetry, infra-red (IR)        transmission through a lead glass viewing coupler, radio        frequency (RF) telemetry, etc. Prior operations in fume hoods        using hot cells are not capable of receiving an RF signal into a        sealed unit because of radiation shielding. This remote control        unit eliminates the risk of the chemist receiving “finger doses”        of radiation during the syntheses process.    -   e) Utilization of remote sensing, including video, that limits        the actuation range of the robotic arms for safety purposes as        well as providing to the chemist a remote visual monitoring        capability. This enables the chemist to either utilize the        remote control at a position distant from the fume hood/hot cell        (e.g., out of viewing range) or to integrate the control        functions of the “remote control” with a more visual environment        outside of the hotcell (e.g. video monitors, personal computers,        etc.).    -   f) A “Macro” record function, which allows the user to manually        execute a synthesis step with the unit, wherein the unit        accurately records each actuation, washing, SPE, sequential        reaction, etc. as a function of time. This synthesis can then be        saved as a user preference or transmitted/shared with other        users of a compatible apparatus elsewhere.    -   g) The functionality of the robotic control unit also includes        “memo record” capabilities to include video, instrumental        measurement of operating parameters (temperature, pressure,        etc.) and voice annotations of a particular temporal event as a        way of capturing expertise or research experience for laboratory        notes or for building training/educational material into macros.

The collection of plug-and-play modules can be combined in differentconfigurations to perform different syntheses. The modules areoperatable in a stand-alone fashion (for integration with manual setupsor other automated systems) or integrated into a central “master”control program.

A first embodiment of a device incorporating features of the inventionas shown in FIGS. 1-4 comprises a remote controlled assembly 10, whichincludes robotic arms (two shown) 12,14 that can be moved up and downand forward and backward under remote control in order to performpressurized reactions, solvent evaporation and product transfer (e.g. toanother SPE unit).

The design incorporates a reactor vessel 16 sometimes referred to as areaction vial 16 mounted on a fixed or removable arm 18. Mounted on thefirst robotic arm 12 is a silicone or Teflon septum or plug 20 orsimilar sealing unit capable of providing a tight seal for high pressurereactions. Also mounted on the first robotic arm 12 is a second plug 24with tubular flow conduits 26 to allow delivery of liquid or gaseousreactants, the application of an inert atmosphere, and removal ofsolvents or end products. The assembly of components also allows for theapplication of pressure or vacuum to vessels from an external gas supplyif desirable. The first robotic arm 12 can be moved such that thereaction vial 16 is open, or is sealed by one of the plugs 20 or 24. Theheating block or oil bath 22 is mounted on the second robotic arm 14 sothat it can be moved to within a desired distance from the reactorvessel 16 for very accurate computerized temperature control. Otheraspects of the reaction system and the solid phase extraction (SPE)components are described below and illustrated in the other figures. Itshould be noted that the SPE components can be mounted on a rack orplatform of the remote controlled assembly 10 for convenience and toreduce clutter, or can be a separate, standalone module.

The embodiments contemplate that the device can either be drivenmanually or remotely, where each axis of motion of the robotic arms12,14 (including the heating unit) may be fractionally adjusted.Alternatively, through the use of limiter devices and specific sensorycomponents, not shown, (e.g., light beams, microswitches, proximitysensors and integrated pressure/temperature transducers) the assemblyand individual components can be controlled using a computer interfaceto perform specific operational command functions, such as “heatreaction vessel for 3 minutes at 120° C.”.

FIGS. 1-4 are schematic and functional representation of the roboticsynthesis assembly with the modules in different functional operationalmodes, allowing the reactants to be added to the reaction vessel and thevessel to be sealed so that the reaction can be performed at desiredtemperatures, combinations of temperatures or temperature ranges andpressures for desired periods of time. FIG. 1 shows the reaction module10 prior to implementation of a reaction procedure. FIG. 2 shows apressurized, closed reaction vessel 16 with the second robot arm 14 andheater block 22 raised to heat the vessel and the first robot arm 12lowered with the plug 20 sealing the vessel. FIG. 3 shows a subsequentstage, wherein the sealing plug 20 has been replaced by the second plug24 for performing, using the flow conduit 26, a solvent evaporationprocedure at the completion of the reaction, which can still be underheating, if required. The vessel 16 can then be remotely opened forproduct transfer or the contents of the vessel can be flushed and/orwashed out of the vessel 16 through the same tubular conduit 26 as shownin FIG. 4.

In a preferred embodiment elevated pressure is generated in the reactionvessel as a result of heating and vaporizing a solvent or other liquidplaced with the reactants in the vessel instead of pressurizing from anexternal source. It has been found that high pressures developed in thereaction vessel using an external source can create unexpected problems.For example, referring to FIG. 5, high pressures can be applied to thereaction vessel using nitrogen gas fed through the N₂ manifold 30.However, due to pressure limitations of the valves 70 and sealinglimitations of the inlet lines in the stopper assembly the reactionvessel may tend to leak or the reactants in the reaction vessel can bepushed into the tubes 36 extending into the reaction mixture. This mayresult in inadequate heating or mixing of the reactants,cross-contamination with reagents added at later times, or loss ofreactants and leakage of radioactive materials to the surrounding area.Commercially available reaction equipment are generally limited to ˜50psi pressures and any attempt to increase the pressures beyond thatleads to the problems described above. In the modular reaction assemblydescribed herein, pressurizing of the reaction vessel is preferably notpressurized using an external gas source so as to avoid these problems.Instead, pressure inside the reaction vessel 16 is generated fromvaporization of the materials within the vessel by controlling thetemperature of the vessel. To prevent leaking or excessive loss ofsolvent the reaction vessel is sealed with silicone or Teflon stoppers.These easily hold high pressures, and no tubes extend into the vesselduring the reaction. In that manner pressures of >100 psi can begenerated and maintained without problem and without loss of the solventor reactants. After the reaction is completed, the reaction vessel isopened from a remote location and appropriate tubings are applied to thevessel and placed into the solution in the reaction vessel for transferout using vacuum or pressure. All these operations can be accomplishedremotely without risk of radiation exposure to the operator.

FIG. 5 is a schematic diagram of a single reactor vial 16 showing theconnections to reagent reservoirs 28, pressurized gas in the N₂ manifold30, prior reactor feed 32 and wash, waste, and product recovery 34. Eachof the feed and withdrawal lines 36 entering the reactor vial 16 arerepresented by the tubular flow conduit 26 in the prior figures. (Forease of visualization, the plug 24 is not shown in FIG. 5). The valves70 connect to each source of feed material.

FIG. 6 is a schematic diagram of a single Solid Phase Extraction (SPE)unit 38 for use to recover the reaction products from a reaction modulesuch as reaction module 10. It should be noted that the SPE can eitherbe a single step purification unit 38 or, as shown in FIG. 7, amulti-step separation system 40. Typical components of the SPE are aneluent chamber 37 and a separation cartridge 39 providing a waste stream41 and a purified product stream or product collection chamber 43. Theaddition of a reservoir of wash solution to wash the cartridge aftertrapping, and the addition of a cartridge regeneration/activationsubsystem are also contemplated.

A unique advantage of the modular system described herein is that it canbe infinitely scaleable. An arbitrary number of reaction vessels eachwith their own controllers and telemetry can be “stacked” together toprovide an n-stage reaction with SPE stages inbetween. This concept isillustrated in FIG. 8 which shows multiple individual remote controlledreactor assemblies 10 in series with single or multistage separationmodules 40, 42 between the reactor assemblies 10 to provide an N-stepreaction module 42. Fluid/reagent lines are modular and neatlyinterfaced to the units for low clutter. Each unit, as well as theassembly, can be preprogrammed or manually interfaced through a controlunit 44 with a computer 46 to control and monitor the process.

If the modular robotic synthesis device is to be used in either a fumehood (with some degree of “retro fit” for low-dose radiation handling)or in a hot cell (for larger doses), different control line adaptationsmay be required. However, the system enables the operator to performsophisticated experiments and material production without radiationexposure. In many instances, the control may be facilitated directly bya cable running through the hot cell/fume hood door or access ports.However, in cases where the radiation shielding is completely sealed,telemetry (not shown) may also be performed, for example using IR/LASERtransmission through a lead glass viewing port. In general, telemetrymay be facilitated by cable, IR/LASER communications (includingvisible/line of site), RF, including techniques such as conventional MHzremote control (e.g. 27 MHz), BlueTooth, Cellular, WiFi and otherproprietary formats. Where IR/LASER communication is performed throughthe lead-glass viewing port of a hot cell, it may also be desired toprovide an optical coupler mounted on each side of the glass.

The controller assembly may be configured in various standardarrangements including a dedicated control box with various levers andswitches to intuitively control the radiochemistry synthesis systemmodules or may take the form of a touch screen or touch tablet PC thathas an instrumentation interface relevant to the configuration of therobotics or the configuration of the chemical operations, or other meansused for remote control of equipment. Preferred aspects of the controlsystem include:

a) Limit sensors to ensure that the device operates at safe temperature,pressure, stirring speeds, and other operating parameters, and thatranges and component movement are not exceeded, thus streamlining usersettings/actions.

b) Real-time data acquisition of all the motions and operatingconditions logged against a time stamp for the purpose of “recording”procedural actions of “macro” and “memo” functions for repeating andduplicating the reaction procedure.

c) An “emergency” stop, which de-pressurizes and/or lowers the reactiontemperatures and conditions and stops the robotic units, in case of amalfunction.

d) Temperature, pressure, radioactivity, time, etc. sensors foraccurately recording the conditions within each reaction vessel.

e) “Macro” functions, for recording a series of reaction events inindividual and an N-module radiochemical synthesis system to establish“presets” so that the procedure can be re-run without user intervention.

f) “Memo” functions, so that the user can create a “snapshot” ofreaction conditions (sensor input), including a video record and arecord of voice memo/text comments, the memos being stored as part of alaboratory annotation system and distributed along with a “Macro”, forexample, for educational/training purposes and to aid in repeatingprocedures and reactions.

Aside from the sensor data which captures the reaction information andthe streamlined utilization of modular remote control assemblies 10, itis also contemplated that the system includes a digital capture of videoinformation. A small video camera (not shown) can be focused on thereaction vessel or vial 16. A “ceiling” view camera (not shown) forobserving each robotic arm 12, 14 as well additional video cameras (notshown) for viewing other components of the system can also be included.This enables the system operator to be physically located at a distantfrom the apparatus. This also enables:

a) Routine functions to be performed (e.g. pre-recorded macros) whilebeing remotely monitored

b) Several units running different production activities to be placedwithin a single hot cell, thus optimizing production space

c) “Tele-chemistry” applications, so that several users at differentlocations can collaborate for an experiment, or individuals at a secondlocation can observe.

d) “Master/slave” operations can be conducted whereby an operatorconducts an experiment at a first location and a slave unit at a secondlocation performs a duplicate of the first operation. This can provideadded production capabilities, confirm reproducibility and be used intraining scenarios.

Operable module systems have been constructed and have been used tosynthesize numerous compounds on a routine basis. Using the apparatusand systems described above, examples of the synthesis of severalradioactive labeled materials and the operating conditions for each aredescribed below following the more detailed descriptions of the severaldifferent modules

Examples of additional modules incorporating features of or usable inthe invention described below include but are not limited to a reagentstorage/delivery module (RDM), cartridge purification module (CPM), highpressure robotic reaction module (PRM), microwave reaction module (MRM),external QC/Analysis/Purification Interface Module (APIM) aliquottingmodule (AM), drying module (DM), concentration module (CM), radiationcounting module (RCM) and a capillary reactor module (CRM).

Reagent Storage/Delivery Module (RDM)

The RDM module 100 contains multiple fluid storage reservoirs 102 thatcan accept reagents during the setup process, prior to the introductionof radioactivity into the system. These reagents are stored until neededduring the synthesis, and then delivered by remote control to the RDMoutput 104. In some embodiments, the RDM includes provisions for certainreagents (e.g. unstable, or air-sensitive reagents) to be loaded intothe module just prior to their use in the synthesis. For example, tubingconnected to the loading port of the RDM can extend outside of theradiation-shielded environment to permit injection of freshly-preparedreagents for immediately addition to the reaction vial.

In a preferred configuration, valves 70 are mounted on a reagentmanifold 106 to minimize the number of fittings and tubing, whichimproves reliability and reduces problems due to human error. A firstembodiment of an output configuration of the RDM has all of thereservoirs 102 connect to a common output channel (FIG. 9). FIG. 9 showsan RDM 100 with eight reservoirs 102 connected through valves 70 to adelivery manifold 106. Connected to each reservoir 102, through thevalve 70 is a reactant feed 108 for the required reactant, so thereservoirs can be filled with the appropriate quantity of reactant foreach reaction. Connected to the top of each reservoir 106 is apressurized gas feed 110 to drive the reservoir contents into themanifold 106. One skilled in the art will recognize that the reservoir'scontents can be delivered by other means such as feed pumps or driversfor plungers in each reservoir, for example a syringe pump system. Allof the valves are remotely controlled for delivery of the appropriatereactants at the appropriate time to the reaction vial 16.

In another embodiment such as shown in FIG. 5 each reservoir has anindependent output port connected to the vial 16. The needs of aparticular radiosynthesis process dictates which one is required, forexample if two reagents are incompatible and carryover is important thesecond embodiment is preferred. Alternatively, a hybrid configurationwith some reagent reservoirs connected to a shared channel and someconnected to individual output ports can be used.

In one embodiment of the RDM, eight external reservoirs are connect byLuer® fittings on the top of the manifold. These vessels can be off theshelf reservoirs made of plastic, glass, etc. and typically have volumesranging from a few mL to 25 mL. Reagents are delivered by gas pressurein the headspace above the reagent. This is controlled via 3-waysolenoid valves that can pressurize the headspace or vent the pressure.The use of inert gases in pressure and vent systems is contemplated forprocesses where reagents are sensitive to air, moisture, etc. For easeof operation the volume can be pre-measured into each reservoir and theentire volume delivered.

In a second embodiment multiple reservoirs can be machined directly intothe manifold. This eliminates the majority of the fittings and tubingneeded. In this embodiment, the maximum reagent volume was selected tobe 3 mL, sufficient for the vast majority of radiosyntheses. Thisembodiment was designed to facilitate automated cleaning after use.Cleaning should be performed immediately after synthesis to avoid dryingof feed solutions that can create particles (e.g. salts) that impedeproper future valve function. Cleaning solutions can be introduced viathe gas pressure system, can be added through an additional fillingport, or can be loaded from the module output.

In a further embodiment the manifold is replaced with a microfluidicchip having integrated microvalves for greatly reduced losses,dead-volume, and carryover. Storage volumes could also be much smaller.

In an embodiment with eight reagent storage sites the reagents aredispensed through a common fluid manifold 106 to a specified location.The RDM includes inert check valves (Bio-Chem, New Jersey) for reagentloading, inert three-way valves (SMC, Japan) for switching between thecommon inlet and reagent loading, and a machined manifold fabricatedfrom Ultem® polyether imide (SABIC Innovative Plastics, Pittsfield,Mass.) to provide the common outlet (FIG. 9). Storage reservoirs used inthe RDM can comprise commercially-available empty, disposable SPEcartridges, or glass reagent vessels. Each reservoir attaches to theUltem® manifold through a Luer-Lok® fitting. The initial reagent loadingis also accomplished through a Luer-Lok connector that is connected tothe check valve. The reservoirs are pressurized at the capped end todispense the stored reagents.

Functional testing of the RDM was undertaken using common reagents usedin radiochemistry (Acetonitrile, DMSO, etc.). Reagents were added anddispensed in a repeated fashion to determine loading and dispensingefficiency. The efficiency was determined by testing performed withF-18, to measure the amount of liquid not dispensed or left behind ineach component of the RDM (fittings, reservoir, etc.). Each componentwas placed in a dose calibrator to determine the amount of F-18 nottransferred through the manifold. Each measurement was compared to thefinal dispensed amount in order to track all of the F-18 activity.Carryover was measured by dispensing radioactive solution from oneposition, then a non-radioactive solution from a second position, andmeasuring the amount of radioactivity in the fluid from the seconddispensing operation.

Cartridge Purification Module (CPM)

The CPM 130 shown in FIG. 12 is designed to perform a variety ofcartridge purifications, generally consisting of the trapping thedesired product on a solid support, washing away contaminants and theneluting the desired product from the solid support. Alternatively, theCPM can also perform ‘filtering’ operations. The CPM includes reagentstorage reservoirs 132 for diluents (to dilute the sample before passingthrough the cartridge, e.g. 1:10 or 1:20 in water), washing, and eluentsolutions along with controlled means to feed these reservoirs. The CPMalso includes an inlet port 133 for the unpurified sample, a cartridge136 (e.g. pre-activated), a waste output 140, a mixing reservoir 138 anda collection output 139 that is connected to the next module. Contentsof reagent storage reservoirs can be directed to flow through thecartridge with appropriate configuration of valves and activation ofpumps (or gas pressure). Fluid flowing through the cartridge can becollected or directed to waste by appropriate configuration of valves.

In one embodiment valves are included in a manifold to minimize thenumber of fittings and tubing. The dilution, wash, and eluent reservoirsare connected via a valve to a common channel which is connected to thepurification cartridge 136. The output of the cartridge is connected tovalves for selection as to whether the flow goes to the waste 140 orcollection (output) port 139.

A second embodiment has two optional features: (i) an additional reagentreservoir of activating solution to perform cartridge activation justprior to the separation, and (ii) an empty reservoir 138 at thecartridge outlet into which the eluate can be loaded and mixed (e.g. bybubbling) to eliminate the concentration gradient as the sample comesoff the cartridge. This may be useful depending on the downstreammodule. For example, a capillary reaction module would have betterperformance if the entire solution has a uniform concentration.

Alternative designs use external reservoirs or embedded reservoirs asdescribed above for the RDM, as well as a microfluidic implementationcan be used.

FIG. 12 shows one embodiment of the CPM. The chemical being purified isfed to product reservoir 134 initially containing a diluents (e.g.water). After delivery of the chemical by gas pressure, additional flowof gas causes bubbling that mixes the chemical with the diluent. Thediluted chemical is then caused to flow through the purificationcartridge 136. In purifications where the desired compound is trapped,the cartridge is then treated with wash solution from a reagentreservoir 132, and partially dried to remove the majority of liquid. Theuntrapped chemical solution and wash solution are directed to the wasteport 140 after passing through the cartridge. The product is released byflowing an eluant (also from a reservoir 132) to transfer the purifiedproduct to the output 139, where the product is collected or isdelivered to the next module in the radiosynthesis system.

The CPM is similar to the RDM in its basic design. However, the fluidpath is slightly different as a purification cartridge 136 is placed inline with the common outlet. Reagent storage reservoirs 132 areconnected through Luer-Lok® ports for easy assembly and removal. Thecartridge 136 is also assembled in the same manner. Reagents forelution, wash, or other purposes can be loaded and delivered throughchemically-inert 3 way valves 70. Loading of the reservoirs 132 isaccomplished by feeding through a chemically inert check valve 70.Delivery of the reagents is accomplished from a pressure source 110supplied to each reservoir. Functional testing was performed aspreviously described for the RDM. Solutions containing F-18 were passedthrough all of the functional components to identify the overall loss ofactivity.

High-Pressure Robotic Reaction Module (PRM)

The main components of the PRM 150, shown in FIGS. 13-17 are a vial 16in which the reaction occurs and a heat transfer unit 22, also referredto as a reaction vessel or heater/reaction vessel. The PRM has an inletport to accept the production solution from the previous module (e.g. apurification module or a reaction module), if one exists, and an outletport connected to the next module in the system.

This PRM is designed to perform high-pressure reactions (typicallyduring superheated conditions when the reaction mixture is heated farabove the solvent boiling point). A reaction “step” may consist ofseveral processes: adding reagents to the vial and mixing, heating(often under sealed conditions to avoid evaporation), and transfer ofproduct out to the next module. Evaporation of solvent before and/orafter the chemical reaction may also be performed. The transfer stepusually requires a dip tube 124 present during the reaction or aretractable tubing/needle (a removeable dip tube 124) that can beintroduced later into the vial 16. However, fixed dip tubes 124 areundesirable when high-pressures are used due to problems describedpreviously, and retractable needles or tubing have reliability concerns.

Commercially available liquid valves can rarely exceed pressures ofabout 50 psi, unless high-pressure “rotary” valves are used. In apreferred embodiment the valves are isolated from the reaction pressureby using robotics to move the vial 16 into a “sealed” configuration(with no paths in fluid connection with valves) during the hightemperature reactions, and then move it into “ported” configurationswhen fluid transfers in and out are needed. In the sealed configurationno valve is exposed to the pressure inside the reaction vial 16; onlythe sealing mechanism is exposed to elevated pressure. As a result, theabove problems are avoided.

In a first embodiment of this module, shown in FIG. 1-4, the reactionvial 16 remains in a fixed position. A heater 22 (e.g. oil bath) can bemoved up to heat the reaction vial 16 and back down to cool the vial 16(e.g. air cooling). Two or more stoppers 20, 24 can be roboticallypositioned (e.g. a 2-axis motion system) to seal with the vial toperform the various operations. The first stopper 20 is typically a plugor cap, while the other stoppers 24 typically have tubing 26 connectedto them to introduce reagents, perform mixing (bubbling), and to drawproduct out of the vial 16 after the reaction.

In a second embodiment shown in FIG. 13, the heater/reaction vessel 22moves so the fluidic interfaces remain stationary. This designaccommodates fluidic interfaces which may include several pieces oftubing 158 connected to other modules in the system. Motion of theseparts would create the potential for this tubing to become tangled,detached, or even damaged. To avoid the risk of failure due to thesefactors, it is preferred to move the heater/reaction vessel 22 instead.The vial 16 is mounted within a thermally-conductive (e.g. metal) heattransfer block so the temperature of the vessel can be controlled at alltimes via active heating (e.g. resistive) and cooling (e.g. air, coolgas, or liquid). A temperature sensor/controller 160 monitors thetemperature inside the metal heater block (calibrated to the liquidtemperature inside the vial) to perform closed loop feedback control ofthe temperature via an integrated temperature controller. Other means ofdelivering energy into the reaction vial are contemplated, includingthermoelectric heating or cooling units, microwaves, heat lamps or laserlight. The vessel 22 and vial 16 move up under control of a motioncontroller 162 which controls a vertical actuator 164 and a horizontalactuator 166 to seal to an interface on a stopper 152, 154, 156 or downto clear any tubing or wiring prior to lateral motion. Lateral motionaligns the vial 16 beneath one of at least 2 different stoppers 152,154, 156 with fluidic interfaces. In a particular embodiment shown inFIGS. 13-17 the PRM has one axis of lateral motion with 3 positions. Thefluidic interfaces provide (i) a sealed surface (for sealed reactions)such as shown in FIG. 17, (ii) tubing 158 for adding reagents such asshown in FIG. 14, (iii) tubing 158 for evaporating solvents, and (iv)tubing 158 and dip tube 124 for transferring product out of the vesselthrough the output stopper 156 (see FIGS. 14-16). Port and tubingconnections and stopper order can be configured for the particular needsof a synthesis. FIG. 14 depicts a typical arrangement: stopper 142 isused for adding reagents and performing solvent evaporations, stopper152 is used for sealed reactions, and stopper 156 is used for producttransfer out of the reaction vial 16.

Sealing is achieved by pushing the top rim of the vial 16 up against aflat elastomeric or plastic gasket layer, typically an insert materialsuch as Viton®, silicone, or Viton® with a protective Teflon® (FEP) filmfor particularly harsh reagents on the stoppers 152, 154, 156. Forexample, the stopper can be constructed from rigid materials (e.g.plastics such as PEEK or Ultem, or metal) with a gasket layer on thebottom surface. The stoppers are installed securely so they remain inplace while the vial is pushed up against the bottom surface. In aspecific embodiment of the present invention, pneumatic force is used tocontrol the sealing force accurately, despite mechanical variations inthe vial dimensions, gasket thicknesses, or mechanical tolerances of thesystem itself. A stepper motor is used to move the vial among the 3fluid interface positions aligned along a single axis. In addition to alinear array, other configurations of fluid interfaces are contemplated,including two-dimensional arrays, or circular arrays (e.g. carousels).Configurations with 2, 3 or more than 3 fluid interface positions arealso envisioned, depending on the needs of the chemical process. Meansof positioning other than pneumatic actuation or stepper motors arepossible, including hydraulics, servo motors, etc. FIGS. 14-17illustrate the sequence of motions as the reaction vial 16 is moved fromfluidic interface 154 in position 1 to the fluidic interface 152 inposition 2.

The robotic system also includes several additional controls: (i) motioncontroller 162 for robotic motion of vial 16; (ii) valves 70 to controlone or more ports of the fluid interface which is easier to vent (topermit filling of reaction vial 16 with reagents) or connected to vacuum(for evaporation), (iii) stir bar actuator (not shown), (iv) valves 70to connect differential pressure to transfer fluid out of the vial 16(positive pressure or vacuum).

As described above, a specific embodiment of the PRM 150 utilizesrobotic control through three stations to facilitate functions requiredfor a chemical reaction. The core component of the PRM 150 is atraditional v-shaped vial or vessel 16 in which all of theradiochemistry processes take place. The vial 16 is placed inside of analuminum heater 22 fixed to a rigid platform. The platform can movevertically using two pneumatically actuated cylinders 164 (SMC, Japan).A stepper motor (Anaheim Automation, CA) drives the platformhorizontally (the horizontal actuator 166). The reactor platform andcylinders are fixed to a seat that moves with the lead screw connectedto the motor. The horizontal motion moves the vial 16 between threestations. Each station is defined by a lid or stopper 152, 154, 156 thatthe reactor platform seals against, using the vertical motion of thepneumatic cylinders. The stopper design is dependent on the functionrequired at each station. For performing sealed reactions, a stopper 152comprising an FEP protective sheet is placed on a viton gasket toprovide a chemically inert surface at volatile conditions. At otherstations, stoppers 154, 156 provide inlets and outlets for the additionor removal of reagents, the supply of heated air, or the removal ofvapor through a vacuum line.

One of the advantages of the PRM 150 over traditional oil-bath/vialreactions is the ability to achieve high pressures and temperatures byincorporating pneumatic cylinders to seal the vial against a chemicallyinert gasket. The PRM 150 utilizes an actively heated and cooledaluminum reactor block (the heater 22). Miniature heater cartridges(Watlow, Mo.) are fixed within the reactor block. Vents machined intothe aluminum are supplied with regulated air pressure, cooled gas, orcooled liquid for active cooling. Temperature control is accomplishedthrough a k-type thermocouple embedded in the reactor, which providesfeedback to the controller 160 (Omega, Conn.). Calibration oftemperature within the vial 16 was accomplished by placing a secondthermocouple through a brass Swagelok fitting embedded in the gasketcovered lid used for sealing. Temperature profiles were then acquired todetermine the lag time of the vial 16 temperature compared to thethermocouple measurement embedded in the reactor. Two tests wereperformed on the gasket seal: increasing pressure by direct injection ofcompressed air (up to 200 psi) into the reaction vial, and internalpressure generation by superheating volatile organic solvents(acetonitrile or dichloroethane up to 200° C.) in the sealed reactionposition. The tests were each performed at least 3 times, for 1 hour.Vial contents were measured before and after tests to determine anyevaporation losses due to poor sealing.

FIGS. 10 and 11 are a schematic representation of liquid transfersubsystems 120 that moves product solution from the reaction vial 16 tothe next module in the system (for example, a purification module). Inone embodiment, shown in FIG. 10, a valve 70 pressurizes the headspaceabove the liquid and forces it into the dip tube connected to the nextmodule in the system. In another embodiment, shown in FIG. 11, a pump112 draws the solution out of the reaction vial 16. A vent permits airor inert gas to replace the liquid removed from the vial.

Microwave Reaction Module (MRM)

The MRM 170 shown in FIG. 18 consists of a microwave cavity 172 (such asa miniature cavity provided by CEM Corporation) that accepts a reactionvial 16. The cavity 172 has a control unit 174 that may includemicrowave power control, temperature control, cooling gas, temperaturemonitoring, and stir bar control.

The vial 16 has a multi-port adapter interface (“lid”) 176 (for example,PEEK with Kalrez® or Viton® seal) with ports for tubing 158 and tubingconnector. These ports can be connected to other modules, e.g. a reagentdelivery module 100, a vacuum evaporation system (described below), thepressurized transfer system as described above, and the downstreammodule (e.g. purification module 130) from the dip tube 124. In additionto control of the microwave reaction itself, this module has auxillarycontrols 120, for example, for venting the vial as reagents are added,applying vacuum and/or a gas stream for evaporations, or applyingdifferent pressure to transfer product out of the reactor. The MRM hasan input port for the reaction product of the previous module (if oneexists), and an output port connected to the next module in the system(if one exists).

No commercial synthesizer contains a microwave energy delivery system.This module permits this capability to be integrated into any process(e.g. existing manual setup, or a modified automated system).Furthermore, this platform permits comparison of a microwave reactorwith a conventional reactor without changing any other parts of thesystem. One skilled in the art will recognize that delivering microwaveenergy to the reactants in the vial 16 in place of conventional heatingmay vary the reaction parameters, particularly reaction time, and theratio of end products as the microwave energy couples directly with thereactants and may not in fact directly heat the solvents in which thereactants are delivered to the vial 16.

External QC/Analysis/Purification Interface Module (APIM)

The APIM 180 shown schematically in FIG. 19, provides an interfacebetween the modular platform and an external system for QC, analysis ofreaction mixtures, or purification (e.g. HPLC).

The APIM has an input 182 for the reaction mixture and an output 184connected to the next module in the system (if one exists). In oneembodiment, this module uses a rotary injection valve 186 to capture acertain volume in an injection loop 188 (a small volume foranalytical/sampling purposes, or the entire volume for preparativepurification purposes). The valve 186 is switched from input to outputto inject this volume into the external system. Any remaining volume(above the injection loop volume) is passed through to the next modulein sequence. The module includes a sample analyzer 190 and a pump 192for moving the withdrawn sample. There are many types of QC, analysis,and purifications that can be performed. For radio-HPLC, the pump 192could be an HPLC pump and the sample analyzer 190 could be an HPLCcolumn and detector system. For radio-TLC, NMR, GC-MS or otheranalytical systems, the pump 192 could simply be pressurized gas to pushthe injection loop volume into a collection vial followed by manualtransfer to the instrument. Full integration with an automated injectionsystem is also contemplated.

This sample collection and transfer function of this module can beimplemented in many ways, e.g. with solenoid valves, or a microfluidicchip if connected to a relatively low-pressure QC/analysis/purificationsystem.

Aliquotting Module (AM)

Dividing a product into multiple doses or to perform reactiondevelopment often required splitting a radioactive sample into N equal(or non-equal) volumes for further processing.

The AM 200, shown in FIG. 20 accepts a sample or stream and divides itamong a programmable set of outlets 202. In one embodiment, a multi-portselection valve 204 is used. The volume directed to the selected outputis metered using a metering pump 206 (e.g. piezo-based micro-pump,syringe pump, etc.), or by repetitive filling and flushing of afixed-volume injection loop. Both approaches can be adapted to providethe capability to deliver a distinct (programmable) volume to eachoutput.

Alternatively a microfluidic implementation of this module that willhave integrated valves to eliminate dead-volumes and reduce loss andcarryover can be used.

F-18 Drying Module

The drying module 210 shown in FIG. 21 performs drying and phasetransfer of [¹⁸F] fluoride ion from its original form in dilute¹⁸O-water from the cyclotron into a mixture of acetonitrile (or otherdry, organic solvent) with K.2.2.2 (Kryptofix) a phase transfer agent,and K₂CO₃. Other phase transfer agents could alternatively be used. Themodule has an input for the aqueous [¹⁸F]fluoride solution and an outputfor the dried complex in organic solvent.

In one embodiment, the module uses an evaporative process, firstevaporating water, then azeotropically removing residual water byadditions and evaporations of dry acetonitrile. Finally, the[¹⁸F]fluoride is formulated into the final solvent and delivered to thenext module. In one embodiment, evaporation is achieved by heating thevial and additionally injecting a stream of heated gas and applyingvacuum. The auxiliary controller that performs these functions has alsobeen described above in the context of the pressurized reaction module,microwave reaction module, etc. and could be considered a distinctmodule. Typical components are a temperature controller 212 connected toa gas heater 214 as well as a vacuum pump 216, the operation of eachcontrolled by a module controller 218. Methods contemplated of heatingthe vial content include the use of a heating block, the use ofmicrowave energy, etc.

Alternatively, microfluidic replacements for this module based onmicrofluidic evaporation or electrochemical trapping and releasetechniques can be used.

More generally, the hardware of this module can be used as asolvent-exchange module (when solvent must be replaced with anothersolvent during the synthesis), or simply an evaporation module.

Concentration Module (CM)

The drying module 210 can also function to concentrate the productsample. This module has an input for the solution to be concentrated,and an output for the concentrated solution. After purifications, suchas by solid-phase extraction (SPE) or HPLC, the volume of the sample maybe increased to several mL or 10's of mL. This volume is too large toperform later chemical reaction steps. Thus, the volume must first bereduced. In some cases, the final product (at the end of the synthesis)must be reduced in volume to meet injection requirements for the patientor research animal.

In one embodiment, the CM such as shown in FIG. 21 uses vacuum-assistedevaporation to remove solvent to concentrate a sample. The sample isloaded into a vial 16 that functions as an evaporation reservoir. It isthen heated (temperature determined according to chemistry and stabilityof solute) and vacuum applied to remove solvent until the desired degreeof concentration has been achieved.

Alternatively a microfluidic version of this module, where the solventis evaporated through a gas-permeable membrane, optimizing the tradeoffbetween large surface area (for fast evaporation) and small surface area(for low loss of reaction mixture).

Radiation Counting Module (RCM)

The RCM measures the radioactivity of a liquid sample. Twoimplementation examples are shown in FIGS. 23 and 24. This module has aninlet to accept a sample to be measured, one outlet to transfer thissample to the next module in the system and a radiation detector tomeasure the radioactivity of the sample.

In a first embodiment 220, the sample is loaded into a vial 16 inproximity to a calibrated radiation sensors 222 by a liquid transfersystem 224. A measurement is made and the sample is transferred out.

In a second embodiment 230, the sample flows in a channel 234 throughthe system. A flow rate sensor 232 in combination with a calibratedradiation sensor 222 is monitored to integrate the total radioactivitypassing the detection point and arrive at the total radioactivity. FIG.22 is a schematic of the radiation sensor with controller and readout240.

There are a wide variety of methods than can be used to measure theradioactivity, including but not limited to PMTs and solid-statesensors.

Capillary Reactor Module (CRM)

The CRM 250 shown in FIG. 25 performs reactions in capillaries ormicrochannels. Two or more liquid streams 252 are pumped into a mixer254 and then fed into capillary tubes or channels in a microfluidic chip256 heated by an energy transfer subsystem 258, possibly connected to acontroller 260. The streams can be pumped at different rates, ordifferent ratios to achieve different residence times through thecapillary.

In one embodiment, the pumps are syringe pumps that first load the twoliquid samples into injection loops and then pump these samples at thedesired flow rates into the capillary tubes or microchannels 256.

Commercial radiosynthesizers based on capillary and chip reactionsexist. However, this subsystem is envisioned as a standalone reactionmodule that can be integrated into a multi-module radiochemicalsynthesis platform. It presents the same interface as the PRM or MRM(inlet for product of previous module, inlet for new reagents, andoutlet to the next module) such that these three reactor types areessentially interchangeable.

FIG. 26 shows a schematic representation of a multi-module reactionsystem including a reaction module (PRM) 150, cartridge purificationmodule (CPM) 130, reagent delivery module (RDM) 100 and an auxiliaryreaction subsystem 120 responsible for venting during reagent addition,evaporation, and product transfer after reaction. The reaction vial 16is mounted within a heat transfer block including heating controller 262and a cooling controller 264 such as used in the following examples.

Example 1 [¹⁸F]FAC Synthesis Non-Robotic

To develop a process for the radiosynthesis of [¹⁸F]FAC on the modularradiochemical synthesis system, steps are first optimized by performingmanually with PRM modules as follows. “Manual” means that all thereagents were added directly into the reaction vial 16 manually (byhand) and only the PRM 150 is used to perform the reaction without usingRDM 100 and CPM 130. Operations of the PRM were operated withremote-control units with touch-screen interfaces. The three PRM unitsare identified as #1, #2 and #3.

Fluorination using PRM #1

The first step of [¹⁸F]FAC synthesis is critical to the finalradiochemical yield. The procedure is as follows:

-   -   Load F-18 solution into a vial containing the solution of K₂CO₃        and K₂₂₂ in MeCN: H₂O (95:5)    -   Measure activity    -   Install vial into heat transfer block    -   Use evaporation system to dry to get [K⊂K2.2.2][¹⁸F]F    -   Add precursor (tribenzoyl pentose triflate, ˜10 mg) in MeCN (0.5        mL) into the reaction vial (RV)    -   Seal RV by moving to position 2 (See FIG. 27). Heat RV to        165° C. for 15 min for radiofluorination    -   Cool down RV below 60° C.    -   Check the conversion yield using radio-HPLC (C-18 column, 1        mL/min, MeCN: H₂O (70:30) as mobile phase, UV setting @ 254 nm)        and radio-TLC (SiO₂, acetone: hexanes (30:70) as developing        agent)    -   Pass the reaction mixture through silica cartridge (pretreated        with hexanes)    -   Elute ¹⁸F-1 out using 4×1 mL of EtOAc into a collection vial    -   Measure the activity of ¹⁸F-1 to obtain the radiochemical yield    -   Check the radiochemical purity of ¹⁸F-1 using radio-HPLC and        radio-TLC    -   This procedure is repeated adjusting operating parameters        (temperature, time) until a repeatable/reasonable yield was        obtained.        Bromination and Cytosine Coupling Via PRM #2

These two steps involve two sensitive reagents, i.e. HBr and silylatedprecursor. The procedure is as follows:

-   -   ¹⁸F-1 solution from the above procedure is added to RV of a        second PRM;    -   Evaporate EtOAc elution to dryness at 90° C. with hot air and        under vacuum in Position 1 (See FIG. 28).    -   Measure activity to determine if there is any activity loss due        to evaporation    -   Add 0.1 mL of HBr in acetic acid to RV, handling HBr within dry        box    -   Add 0.4 mL of dichloroethane into RV    -   Seal RV (Position 2) and heat RV at 75° C. for 10 min for        bromination    -   Evaporate HBr/AcOH/dichloroethane at 75° C. to about half of the        original volume    -   Measure activity (to determine if there is any activity loss due        to evaporation)    -   Add 0.7 mL of toluene into RV    -   Evaporate to dryness at 110° C. assisted with hot air and vacuum        in Position 1    -   Measure activity (to determine if there is any activity loss due        to evaporation)    -   Add 50 mg of silylated precursor in 1 mL of dichloroethane into        RV    -   Seal RV (Position 2) and heat RV to 160° C. for 30 min for        coupling    -   Cool down below 60° C.    -   Check the coupling yield using radio-HPLC and radio-TLC    -   Pass the reaction mixture through silica gel cartridge        (pretreated with hexanes)    -   Elute ¹⁸F-3 out using 5×1 ml of MeOH:CH₂Cl₂ (10:90) into a        collection vial    -   Measure the activity of ¹⁸F-3 to check the radiochemical yield    -   Obtain the radiochemical purity of ¹⁸F-3 using radio-HPLC and        radio-TLC    -   This procedure is repeated, tuning reaction conditions and        observing outcomes until a process for repeatable/reasonable        yield is achieved.        Deprotection using PRM #3

The deprotection step also involves a sensitive reagent, sodiummethoxide solution.

-   -   Add ¹⁸F-3 solution prepared above to RV of PRM #3    -   Evaporate elution solution (MeOH:CH₂Cl₂ (10:90)) to complete        dryness at 100° C. assisted with hot air and vacuum in Position        1    -   Measure activity (to determine if there is any activity loss due        to evaporation)    -   Add 0.5 mL of 0.5M sodium methoxide solution into RV    -   Seal RV (Position 2) and heat to 100° C. for 5 min    -   Cool down below 60° C.    -   Add 0.25 mL of 1N HCl into RV    -   Check the reaction yield using radio-HPLC (C-18 column, 1        mL/min, EtOH: 50 mM NH₄OAc in water (10:90) as mobile phase, UV        setting @ 254 nm) and radio-TLC (SiO₂, CH₂Cl₂: MeOH (85:15) as a        developing agent) (See FIG. 29)    -   Evaporate MeOH out of the solution at 100° C. assisted with hot        air and under vacuum in Position 1    -   Measure activity (to determine if there is any activity loss due        to evaporation)    -   Add 3 mL of HPLC mobile phase into RV to dilute the residue    -   Inject into preparative HPLC and collect fractions    -   Measure the activity of ¹⁸F-4 to obtain the radiochemical yield    -   Check the radiochemical purity of ¹⁸F-4 using radio-HPLC and        radio-TLC    -   This procedure is repeated and reaction parameters adjusted        until a repeatable/reasonable yield is obtained.

Example 2 Synthesis of [¹⁸F]FAC in the Integrated System, Including RDM,PRM and CPM

“The integrated ARC-P system” includes three sequential sets of RDM 100,PRM 150 and CPM 130 as shown in FIGS. 30-32. All of the reagents forreaction and separation were added into the reaction vial (RV) 16 bypassing through the RDM 100. The PRM 150 is used to perform evaporationsand heating, and the CPM 130 is used for the cartridge purification. Thepreparation of [¹⁸F] FAC follows the reaction scheme and the descriptiongiven below

Fluorination (Referring to FIG. 30)

-   -   Transfer aqueous F-18 solution containing K₂₂₂/K₂CO₃/MeCN from        chamber 1 (R1) in the RDM manifold into the reaction vial (RV)        to prepare [K⊂K2.2.2][¹⁸F]F    -   Heat RV to 110° C. for 1 min assisted with hot air and vacuum in        Position 1 (1^(st) F-18 drying)    -   Cool down below 60° C.    -   Transfer 1 mL of MeCN from R7 into RV    -   Heat RV to 110° C. for 1 min assisted with hot air and vacuum in        Position 1 (2^(nd) drying)    -   Cool down below 60° C.    -   Transfer 1 mL of MeCN from R8 into RV (3rd    -   Heat RV to 110° C. for 3 min assisted with hot air and vacuum in        Position 1 drying)    -   Heat RV at 110° C. for additional 3 min with vacuum only in        Position 1 (final drying step)    -   Transfer the precursor (tribenzoyl pentose triflates, 10 mg) in        0.5 mL of MeCN from R3 into RV    -   Seal and heat RV to 165° C. for 15 min in Position 2 for        radiofluorination    -   Cool down below 60° C.    -   Check the conversion yield using radio-HPLC (C-18 column, 1        mL/min, MeCN:H₂O (70:30) as mobile phase, UV setting @ 254 nm)        and radio-TLC (SiO₂, acetone:hexanes (30:70) as developing        agent)    -   Pass the reaction mixture through silica gel cartridge        (pretreated with hexanes)    -   Elute ¹⁸F-1 out using 4×1 mL of EtOAc into RV #2    -   Measure the activity of ¹⁸F-1 to obtain the radiochemical yield    -   Check the radiochemical purity of ¹⁸F-1 using radio-HPLC and        radio-TLC

Bromination and Cytosine Coupling (FIG. 31)

-   -   Transfer ¹⁸F-1 solution to RV of PRM #2    -   Evaporate eluate to dryness at 90° C. assisted with hot air and        vacuum in Position 1    -   Measure activity to determine if there is any activity loss due        to evaporation.    -   Add 0.1 mL of HBr in acetic acid to RV #2    -   Transfer 0.4 mL of dichloroethane from R3 into RV #2    -   Seal and heat RV to 75° C. for 10 min in Position 2 for        bromination    -   Evaporate HBr/AcOH/dichloroethane at 75° C. to about half of the        original volume    -   Measure activity to determine if there is any activity loss due        to evaporation    -   Transfer 0.7 mL of toluene from R5 into RV    -   Evaporate to dryness at 115° C. (110° C. inside the vial) with        hot air of 130° C. and under vacuum (−10˜15 in. Hg) in Position        1    -   Measure activity to determine if there is any activity loss due        to evaporation    -   Add 50 mg of silylated precursor in 1 ml of dichloroethane into        RV by passing either through a tubing outside the hotcell        remotely, or via RDM    -   Seal and heat RV to 160° C. for 30 min in Position 2 for        coupling    -   Cool down below 60° C.    -   Check the coupling yield using radio-HPLC and radio-TLC    -   Pass the reaction mixture through silica cartridge pretreated        with hexanes    -   Elute ¹⁸F-3 out using 5×1 mL of MeOH: CH₂Cl₂ (10:90) from R7        into a collection vial or RV #3    -   Measure the activity of ¹⁸F-3 to check the radiochemical yield    -   Check the radiochemical purity of ¹⁸F-3 using radio-HPLC and        radio-TLC

Deprotection (FIG. 32)

-   -   Transfer ¹⁸F-3 solution to RV #3 in PRM #3; Evaporate elution        solution (MeOH:CH₂Cl₂ (10:90)) to complete dryness at 100° C.        assisted with hot air and vacuum in Position 1    -   Measure activity    -   Transfer 0.5 mL of 0.5M sodium methoxide solution into RV #3 by        passing, either through a tubing outside the hotcell remotely,        or via RDM    -   Heat to 100° C. for 5 min in Position 2    -   Cool down below 60° C.    -   Transfer 0.25 mL of 1N HCl from R3 into RV #3    -   Check the deprotection yield using radio-HPLC (C-18 column, 1        mL/min, EtOH: 50 mM NH₄OAc in water (10:90) as mobile phase, UV        setting @ 254 nm) and radio-TLC (SiO₂, CHCl₃: MeOH (10:90) as        developing agent)    -   Evaporate MeOH out of the solution at 100° C. assisted with hot        air and vacuum in Position 1    -   Measure activity    -   Transfer 3 mL of HPLC mobile phase from R5 into RV #3 to dilute        the residue    -   Inject solution in RV #3 into preparative HPLC and collect        fractions    -   Measure the activity of ¹⁸F-4 to check the radiochemical yield    -   Check the radiochemical purity of ¹⁸F-4 using radio-HPLC and        radio-TLC.

Example 3 Preparation ofD-2′-deoxy-2′[¹⁸F]fluoro-β-D-arabinofuranosylcytosine (D-[¹⁸F]FAC)

The preparation of D[¹⁸F] FAC follows the reaction scheme and thedescription given below:

2-O-[(Trifluoromethyl)sulfonyl]-1,3,5-tri-O-benzoyl-α-D-ribofuranose (1)was prepared as reported in the literature (Tann, C. H., Brodfuehrer, P.R., Brundidge, S. P., Sapino, Jr. C., and Howell, H. G.“Fluorocarbohydrates in Synthesis. An Efficient Synthesis of1-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)-5-iodouracil (β-FIAU) and1-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)thymine (β-FMAU)”. J. Org.Chem., 50, pp 3644-3647(1985)) The synthesis of the ¹⁸F-fluoro analog 2was carried out by a modification of the method reported by Mangner etal. (Mangner, T. J., Klecker, R. W., Anderson, L., and Shields, A. F.“Synthesis of 2′-deoxy-2′[¹⁸F]fluoro-β-D-arabinofuranosyl nucleosides,[¹⁸F]FAU, [¹⁸F]FMAU, [¹⁸F]FBAU and [¹⁸F]FIAU, as potential PET agentsfor imaging cellular proliferation”, Nuc. Med. Biol., 50, pp 215-224(2003).

The radiosynthesis was carried out using three of the robotic reactionmodules of FIGS. 1-4. No-carrier-added [¹⁸F]fluoride ion was produced by11 MeV proton bombardment of 98% enriched [¹⁸O]water in a silver targetbody using a RDS-112 cyclotron. The aqueous [¹⁸F]fluoride ion wastreated with a solution of K₂CO₃ (1 mg) and Kryptofix 2.2.2 (10 mg)dissolved in water (0.04 mL) and acetonitrile (0.75 mL) mixture. Thesolution was evaporated at 115° C. with a stream of nitrogen gas. Theresidue was dried by the azeotropic distillation with acetonitrile(3×0.5 mL). A solution of the triflate 1 (10 mg) in 0.7 mL ofacetonitrile was added to the dry residue and the reaction mixture washeated at 165° C. for 15 min in a sealed vessel. The solution was cooledto room temperature and passed through a small cartridge of silica gel.The product was eluted from the cartridge with 5 mL of ethyl acetate.The ethyl acetate solution was evaporated to dryness and 0.1 mL of asolution of 30% HBr in acetic acid was added followed by 0.4 mL ofdichloroethane. This new reaction mixture was heated at 80° C. in asealed vessel for 10 min and the solution was concentrated to ˜50% ofthe initial volume. Toluene (0.7 mL) was then added and the solution wasevaporated at 110° C. to give the bromo compound 3. A freshly prepareddisilyl derivative of cytosine (4, 35 mg) was dissolved in 1 mL ofdichloroethane and added to the bromo compound 3. The condensationreaction was carried out at 160° C. in a sealed vessel for 30 min. Thereaction mixture was cooled to room temperature and then passed througha small column of silica gel. The product was eluted off the columnusing 5 mL of a solution mixture of 10% methanol and 90%dichloromethane. This solution was evaporated to dryness at 100° C. andthen treated with 0.5 mL of a solution of 0.5 M sodium methoxide inmethanol. The reaction mixture was heated at 100° C. for 5 min in asealed vessel. The basic reaction mixture was neutralized with 0.25 mLof 1M HCl in water. This reaction mixture was diluted to a total volumeof 3 mL with a mixture of 1% ethanol and 99% 10 mM ammonium dihydrogenphosphate in water and injected into a semi-preparative HPLC column(Phenomenex Gemini C-18 column; 25 cm×1 cm). The HPLC column was elutedwith a solvent mixture of 1% ethanol and 99% 10 mM ammonium dihydrogenphosphate at a flow rate of 5.0 mL/min. The effluent from the HPLCcolumn was monitored with a 254 nm UV detector followed by a gammaradioactive detector. The chemically and radiochemically pure D-[¹⁸F]FAC that eluted off the column with a retention time of ˜15 min was madeisotonic with normal saline and sterilized by passing through aMillipore filter (0.22 μm) into a sterile multi-dose vial.

Chemical and Radiochemical Quality Control

The chemical and radiochemical purities of D-[¹⁸F] FAC were determinedby an analytical HPLC method using a Phenomenex Luna column (25 cm×0.46cm, 5μ particle size). The column was eluted with 10% ethanol and 90% 50mM ammonium acetate at a flow rate of 1.0 mL/min. The effluent from theHPLC column was passed through a UV detector (λ=254 nm) followed by agamma radioactivity detector. The chemical and radiochemical purities ofD-[¹⁸F] FAC prepared as described above exceeded 99.9% as shown in theanalytical HPLC chromatogram (see FIG. 37).

Analytical HPLC also was used to determine the specific activity ofD-[¹⁸F] FAC. A range of mass vs UV absorption at 254 nm wavelength fornon-radiolabeled D-FAC was determined using the analytical HPLC methoddescribed above and the data set was used to construct a calibrationgraph. Using this calibration graph, the specific activity of D-[¹⁸F]FAC was found to be 1000 Ci/mmol.

Radionuclide Analysis

A calibrated γ-ray spectrometer was used to establish the presence ofthe 511 keV annihilation radiation associated with the decay of ¹⁸Fisotope.

Sterility and Pyrogenicity Tests

D-[¹⁸F] FAC prepared as described above was tested for sterility usingthe standard thioglycollate medium procedure and found to be sterile.

The absence of pyrogens in the D-[¹⁸F] FAC preparation was verified bythe standard Limulus Amebocyte Lysate (LAL) test.

Example 4 Preparation ofL-2′-Deoxy-2′-[¹⁸F]fluoro-β-D-arabinofuranosylcytosine (L-[¹⁸F] FAC)

The preparation of L-[¹⁸F] FAC follows the reaction scheme and thedescription given below:

L-2-O-[(Trifluoromethyl)sulfonyl]-1,3,5-tri-O-benzoyl-α-D-ribofuranose(1) was prepared based on the procedure reported by Tann et al. for thecorresponding D-isomer (Example 1).

The synthesis of the ¹⁸F-fluoro analog 2 was carried out by amodification of the above referenced method reported by Mangner et al.described for the corresponding D-isomer.

The radiosynthesis was carried out using three of the robotic reactionmodules of FIGS. 1-4. No-carrier-added [¹⁸F]fluoride ion was produced by11 MeV proton bombardment of 98% enriched [¹⁸O]water in a silver targetbody using a RDS-112 cyclotron. The aqueous [¹⁸F]fluoride ion wastreated with a solution of K₂CO₃ (1 mg) and Kryptofix 2.2.2 (10 mg)dissolved in water (0.04 mL) and acetonitrile (0.75 mL) mixture. Thesolution was evaporated at 115° C. with a stream of nitrogen gas. Theresidue was dried by the azeotropic distillation with acetonitrile(3×0.5 mL). To the dry residue, a solution of the triflate 1 (10 mg) in0.7 mL of acetonitrile was added and the reaction mixture was heated at165° C. for 15 min in a sealed vessel. The solution was cooled to roomtemperature and passed through a small cartridge of silica gel. Theproduct was eluted from the cartridge with 5 mL of ethyl acetate. Theethyl acetate solution was evaporated to dryness and 0.1 mL of asolution of 30% HBr in acetic acid was added followed by 0.4 mL ofdichloroethane. This new reaction mixture was heated at 80° C. in asealed vessel for 10 min and the solution was concentrated to ˜50% ofthe initial volume. Toluene (0.7 mL) was then added and the solution wasevaporated at 110° C. to give the bromo compound 3. A freshly prepareddisilyl derivative of cytosine (4, 35 mg) was dissolved in 1 mL ofdichloroethane and added to the bromo compound 3. The condensationreaction was carried out at 160° C. in a sealed vessel for 30 min. Thereaction mixture was cooled to room temperature and then passed througha small column of silica gel. The product was eluted off the columnusing 5 mL of a solution mixture of 10% methanol and 90%dichloromethane. This solution was evaporated to dryness at 100° C. andthen treated with 0.5 mL of a solution of 0.5 M sodium methoxide inmethanol. The reaction mixture was heated at 100° C. for 5 min in asealed vessel. The basic reaction mixture was neutralized with 0.25 mLof 1M HCl in water. This reaction mixture was diluted to a total volumeof 3 mL with a mixture of 1% ethanol and 99% 10 mM ammonium dihydrogenphosphate in water and injected into a semi-preparative HPLC column(Phenomenex Gemini C-18 column; 25 cm×1 cm). The HPLC column was elutedwith a solvent mixture of 1% ethanol and 99% 10 mM ammonium dihydrogenphosphate at a flow rate of 5.0 mL/min. The effluent from the HPLCcolumn was monitored with a 254 nm UV detector followed by a gammaradioactive detector. The chemically and radiochemically pure L-[¹⁸F]FAC that eluted off the column with a retention time of ˜15 min was madeisotonic with normal saline and sterilized by passing through aMillipore filter (0.22 μm) into a sterile multi-dose vial.

Chemical and Radiochemical Quality Control

The chemical and radiochemical purities of L-[¹⁸F] FAC were determinedby an analytical HPLC method using a Phenomenex Luna column (25 cm×0.46cm, 5μ particle size). The column was eluted with 10% ethanol and 90% 50mM ammonium acetate at a flow rate of 1.0 mL/min. The effluent from theHPLC column was passed through a UV detector (λ=254 nm) followed by agamma radioactivity detector. The chemical and radiochemical purities ofL-[¹⁸F] FAC prepared as described above exceeded 99.9% as shown in theanalytical HPLC chromatogram (FIG. 38).

Analytical HPLC also was used to determine the specific activity ofL-[¹⁸F] FAC. A range of mass vs UV absorption at 254 nm wavelength fornon-radiolabeled L-FAC was determined using the analytical HPLC methoddescribed above and the data set was used to construct a calibrationgraph. Using this calibration graph, the specific activity of L-[¹⁸F]FAC was found to be >1000 Ci/mmol.

Radionuclide Analysis

A calibrated γ-ray spectrometer was used to establish the presence ofthe 511 keV annihilation radiation associated with the decay of ¹⁸Fisotope.

Sterility and Pyrogenicity Tests

L-[¹⁸F] FAC prepared as described above was tested for sterility usingthe standard thioglycollate medium procedure and found to be sterile.

The absence of pyrogens in the L-[¹⁸F] FAC preparation was verified bythe standard Limulus Amebocyte Lysate (LAL) test.

Insert 0076

Example 5 Preparation ofL-2′-Deoxy-2′-[¹⁸F]fluoro-β-D-arabinofuranosyl-5-methylcytosine (L-[¹⁸F]FMAC)

The preparation of L-[¹⁸F] FMAC follows the reaction scheme and thedescription given below:

L-2-O-[(Trifluoromethyl)sulfonyl]-1,3,5-tri-O-benzoyl-α-D-ribofuranose(1) was prepared based on the procedure reported by Tann et al. for thecorresponding D-isomer (referenced above). The synthesis of the¹⁸F-fluoro analog 2 was also carried by a modification of the methodreported by Mangner et al. The radiosynthesis was carried out usingthree of the robotic reaction modules of FIGS. 1-4. No-carrier-added[¹⁸F]fluoride ion was produced by 11 MeV proton bombardment of 98%enriched [¹⁸O]water in a silver target body using a RDS-112 cyclotron.The aqueous [¹⁸F]fluoride ion was treated with a solution of K₂CO₃ (1mg) and Kryptofix 2.2.2 (10 mg) dissolved in water (0.04 mL) andacetonitrile (0.75 mL) mixture. The solution was evaporated at 115° C.with a stream of nitrogen gas. The residue was dried by the azeotropicdistillation with acetonitrile (3×0.5 mL). To the dry residue, asolution of the triflate 1 (10 mg) in 0.7 mL of acetonitrile was addedand the reaction mixture was heated at 165° C. for 15 min in a sealedvessel. The solution was cooled to room temperature and passed through asmall cartridge of silica gel. The product was eluted from the cartridgewith 5 mL of ethyl acetate. The ethyl acetate solution was evaporated todryness and 0.1 mL of a solution of 30% HBr in acetic acid was addedfollowed by 0.4 mL of dichloroethane. This new reaction mixture washeated at 80° C. in a sealed vessel for 10 min and the solution wasconcentrated to ˜50% of the initial volume. Toluene (0.7 mL) was thenadded and the solution was evaporated at 110° C. to give the bromocompound 3. A freshly prepared disilyl derivative of 5-methylcytosine(4, 35 mg) was dissolved in 1 mL of dichloroethane and added to thebromo compound 3. The condensation reaction was carried out at 160° C.in a sealed vessel for 30 min. The reaction mixture was cooled to roomtemperature and then passed through a small column of silica gel. Theproduct was eluted off the column using 5 mL of a solution mixture of10% methanol and 90% dichloromethane. This solution was evaporated todryness at 100° C. and then treated with 0.5 mL of a solution of 0.5 Msodium methoxide in methanol. The reaction mixture was heated at 100° C.for 5 min in a sealed vessel. The basic reaction mixture was neutralizedwith 0.25 mL of 1M HCl in water. This reaction mixture was diluted to atotal volume of 3 mL with a mixture of 1% ethanol and 99% 10 mM ammoniumdihydrogen phosphate in water and injected into a semi-preparative HPLCcolumn (Phenomenex Gemini C-18 column; 25 cm×1 cm). The HPLC column waseluted with a solvent mixture of 2% ethanol and 98% 10 mM ammoniumdihydrogen phosphate at a flow rate of 5.0 mL/min. The effluent from theHPLC column was monitored with a 254 nm UV detector followed by a gammaradioactive detector. The chemically and radiochemically pure L-[¹⁸F]FMAC that eluted off the column with a retention time of 15 min was madeisotonic with normal saline and sterilized by passing through aMillipore filter (0.22 μm) into a sterile multi-dose vial.

Chemical and Radiochemical Quality Control

The chemical and radiochemical purities of L-[¹⁸F] FMAC were determinedby an analytical HPLC method using a Phenomenex Luna column (25 cm×0.46cm, 5μ particle size). The column was eluted with 10% ethanol and 90% 50mM ammonium acetate at a flow rate of 1.0 mL/min. The effluent from theHPLC column was passed through a UV detector (λ=254 nm) followed by agamma radioactivity detector. The chemical and radiochemical purities ofL-[¹⁸F] FMAC prepared as described above exceeded 99.9% as shown in theenclosed typical analytical HPLC chromatogram (FIG. 39).

Analytical HPLC also was used to determine the specific activity ofL-[¹⁸F] FMAC. A range of mass vs UV absorption at 254 nm wavelength fornon-radiolabeled L-FMAC was determined using the analytical HPLC methoddescribed above and the data set was used to construct a calibrationgraph. Using this calibration graph, the specific activity of L-[¹⁸F]FMAC was found to be >1000 Ci/mmol.

Radionuclide Analysis

A calibrated γ-ray spectrometer was used to establish the presence ofthe 511 keV annihilation radiation associated with the decay of ¹⁸Fisotope.

Sterility and Pyrogenicity Tests

L-[¹⁸F] FMAC prepared as described above was tested for sterility usingthe standard thioglycollate medium procedure and found to be sterile.

The absence of pyrogens in the L-[¹⁸F] FMAC preparation was verified bythe standard Limulus Amebocyte Lysate (LAL) test.

Example 6 Synthesis of ¹⁸F-Clofarabine (¹⁸F-CA)

The preparation of ¹⁸F-CA follows the reaction scheme and thedescription given below:

The trityl protected chloroadenosine derivative 2 was prepared by ageneral procedure previously reported (Pankiewicz, K. W., Krzeminski,J., Cizewaki, L. A., Ren, W.-Y., and Watanabe, K. A. “A Synthesis of9-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)adenine and Hypoxanthine. AnEffect of C3′-Endo to C2′-Endo Conformational Shift on the ReactionCourse of 2′-Hydroxyl Group with DAST” J. Org. Chem., 57, pp 553-559(1992)) 2-chloroadenosine (1) (9.2 mmol), 4-dimethylaminopyridine (9.2mmol) and monomethoxytrityl chloride (32.4 mmol) were placed in a dry250 mL round bottom flask under argon and 80 mL of dry pyridine wasadded. The mixture was stirred at 90° C. for 18 h. Pyridine wasevaporated in rotary evaporator and the last traces of it wereazeotropically removed with toluene. The residue was dissolved indichloromethane and washed with water. The organic layer was dried withNa₂SO₄, filtered and evaporated. The crude product was subjected tosilica gel column chromatography with 25% ethyl acetate in hexane as theeluent to isolate pure hydroxy product 2. The triflate 3 was preparedfrom the corresponding hydroxy derivative 2 as follows: The hydroxycompound 2 (0.1 mmol) was dissolved in 3 mL of dichloromethane underargon and 4-dimethylaminopyridine (0.18 mmol) was added. The solutionwas cooled in an ice bath at 0° C. for 10 min. Triflyl chloride (0.02mL) was then added and the reaction mixture was gradually warmed to roomtemperature and stirred for 3 h. The reaction mixture was diluted with10 mL of dichloromethane and washed with water. The organic layer wasdried with Na₂SO₄. Evaporation of dichloromethane gave an oily residue,which was purified by silica gel column chromatography using 30% ethylacetate in hexane as eluent provided the pure triflate derivative 3.

The radiosynthesis was carried out using three of the robotic reactionmodules of FIGS. 1-4. No-carrier-added [¹⁸F]fluoride ion was produced by11 MeV proton bombardment of 98% enriched [¹⁸O]water in a silver targetbody using a RDS-112 cyclotron. The aqueous [¹⁸F]fluoride ion wastreated with a solution of K₂CO₃ (1 mg) and Kryptofix 2.2.2 (10 mg)dissolved in water (0.04 mL) and acetonitrile (0.75 mL) mixture. Thesolution was evaporated at 115° C. with a stream of nitrogen gas. Theresidue was dried by the azeotropic distillation with acetonitrile(3×0.5 mL). The triflate precursor 3 (10 mg) dissolved in 1 mL ofacetonitrile was added to the dried K¹⁸F/Kryptofix complex and reactedat 125° C. for 25 min in a sealed reaction vessel. The reaction mixturewas cooled to room temperature and passed through a small cartridge ofsilica gel. The cartridge was eluted with 4×2 mL of ethyl acetate. Theethyl acetate was evaporated to dryness and the residue was thendissolved in 0.5 mL of acetonitrile. One mL of 1M HCl was added to theacetonitrile solution and heated at 100° C. for 5 min. The reactionmixture was diluted to a total volume of 3 mL with a solution of 15%ethanol and 85% 25 mM ammonium acetate in water and injected into asemi-preparative HPLC column (Phenomenex Gemini C-18 column; 25×1 cm)and eluted with a mobile phase of 15% ethanol and 85% 25 mM ammoniumacetate in water at a flow rate of 5.0 mL/min. The effluent from thecolumn was monitored with an UV detector (λ=263 nm) and a gammaradioactive detector. The chemically and radiochemically pure¹⁸F-labeled product 4 with retention time between 11 and 13 min isolatedin 10-15% radiochemical yield was made isotonic by dilution with sterilesaline solution which also decreased the concentration of ethanol to<10%. The solution was then sterilized by passing through a Milliporesterilizing filter (0.22 μm) into a sterile multi-dose vial.

Chemical and Radiochemical Quality Control

The chemical and radiochemical purities of ¹⁸F-CA, as synthesized above,were determined by an analytical HPLC method using a Phenomenex Lunacolumn (25 cm×0.46 cm, 5μ particle size). The column was eluted with 15%ethanol and 85% 25 mM ammonium acetate at a flow rate of 1.5 mL/min. Theeffluent from the HPLC column was passed through a UV detector (λ=263nm) followed by a gamma radioactivity detector. The chemical andradiochemical purities of ¹⁸F-CA prepared as described above exceeded97% as shown in the analytical HPLC chromatogram (FIG. 40).

Radionuclide Analysis

A calibrated γ-ray spectrometer was used to establish the presence ofthe 511 keV annihilation radiation associated with the decay of ¹⁸Fisotope.

Sterility and Pyrogenicity Tests

¹⁸F-CA prepared as described above was tested for sterility using thestandard thioglycollate medium procedure and found to be sterile.

The absence of pyrogens in the ¹⁸F-CA preparation was verified by thestandard Limulus Amebocyte Lysate (LAL) test.

The examples above illustrate the utility of the disclosed modularsystems are not intended to limit the scope of the invention and aremerely representative of various capabilities of the system.

For example while the use of sealing plugs for each reaction vessel isillustrated, each reaction vessel may have remotely controlled valvedports built therein and feed materials may be delivered through remotelycontrolled manifolds attached to each vessel. Still further, eachreaction vessel may include remotely controlled heaters and coolersintegral therewith.

Example 7 Preparation of [¹⁸F]-fluoro-4-nitrobenzene ([¹⁸F]FNB) in theIntegrated System, Including RDM, PRM and CPM (FIG. 26)

Cleaning and Drying

5-10 ml of H₂O was loaded into each reservoir in the RDM and the CPM andthen transferring out through the manifold, delivery tubing and transfertubing to waste. Using 5-10 ml of ethanol and acetone, the above processwas repeated twice. Finally, all air valves and liquid valves in the RDMand the CPM and the reservoirs, channels, and tubings were dried.

Preloading of Reagents and Solvents

Non-radioactive reagents were preloaded into the correspondingreservoirs for the RDM and the CPM. The reagent configuration of RDM isas follows, 1,4-dinitrobenzene (DNB, 4 mg) in 0.5 ml of DMSO loaded inreservoir #3, 1 ml of H₂O in reservoir #5, 1 ml of H₂O in reservoir #6and 1 ml of anhydrous MeCN in reservoir #8. Reservoir #2, #4 and #7 areempty. Reservoir #1 is used for loading ¹⁸F ion at a later time. Thereagent configuration of CPM is as follows, 10 ml of H₂O in reservoir C,10 ml of H₂O in reservoir A, 2 ml of methanol in reservoir D. Thepurification cartridge (stara C18, 30 mg) was preconditioned with 10 mlof ethanol and H₂O in the corresponding position of CPM.

Production and Activation of [¹⁸F]Fluoride.

No-carrier-added [¹⁸F]F-ion was obtained from the nuclear reaction¹⁸O(p, n)¹⁸F by irradiation of 97% ¹⁸O-enriched water with an 11 MeVproton beam using RDS-112 cyclotron (Siemens). 50-100 μl (10˜30 μCi) ofaqueous [¹⁸F]F-ion solution in [¹⁸O]H₂O was mixed with 20 mg ofKryptofix₂₂₂ (K₂₂₂), 26 μl of 1M of aqueous K₂CO₃ solution and 1 ml ofanhydrous MeCN, loaded into reservoir #1 in RDM using a syringemanually, then transferred into a reaction V-vial (Wheaton) of the PRMthrough the delivery tubing in RDM. The mixed solution was heated at110° C. for 5 min with hot air blowing and vacuum suction to remove thewater by azeotropic evaporation until dry. After compressed-air coolingdown to room temperature, 1 ml of anhydrous MeCN in reservoir #8 wasdelivered into the. The azeotropic evaporation was repeated once usingthe same condition as above. After a final addition of anhydrous MeCN,reactor heating and vacuum anhydrous K₂₂₂/[¹⁸F]F complex.

Radiosynthesis and Purification of [¹⁸F]FNB.

1,4-Dinitrobenzene solution in DMSO in reservoir #3 of RDM was deliveredinto the reaction vial. The reaction mixture was heated at 145° C. for 8min to perform the radiofluorination of precursor and produce thelabeled product [¹⁸F]FNB. After cooling down to room temperature, 1 mlof H₂O in reservoir #5 was delivered into the vial to dilute thereaction mixture. The diluted mixture was transferred into reservoir Cin the CPM to dilute further. 1 ml of H₂O in reservoir #6 of the RDM wasdelivered into the reaction vial to wash and obtain the residualreaction mixture, then transferred into reservoir C in the CPM. Thediluted reaction mixture was passed through the cartridge and theelution was directed into a waste vial. The cartridge was washed using10 ml of H₂O in reservoir A of the CPM. Finally, [¹⁸F]FNB was eluted outusing 2 ml of MeOH in reservoir D of CPM and collected in the productvial.

Quality Control of [¹⁸F]FNB.

The total activity of [¹⁸F]FNB solution in methanol was measured by dosecalibrator and its radiochemical purity was checked by radio-TLC andradio-HPLC for the purpose of quality control. The radio-HPLC spectrumis shown in FIG. 33. The sample was spotted on a TLC plate, which wasdeveloped in pure CH₂C₁₂ and scanned by radio-TLC scanner. The samplewas also injected into the HPLC to analyze, which was run using MeCN/H₂O(V/V 50/50) with 0.2% TFA as a mobile phase at the flow rate of 1ml/min. The HPLC-System comprises a K2501 UV detector (Knauer),B-FC-1000 radiodetector (Bioscan), 501 pump (Knauer) and Luna column (5m C18(2) 100 A, 250×4.6 mm) (Phenomenex) and Gina box (Raytest) for dataacquisition and interpretation. 254 nm was selected as the UV detectionwavelength.

Example 8 Preparation of 1¹⁸F12-fluoro-2-deoxy-D-glucose ([¹⁸F]FDG) inthe Integrated System, Including RDM, PRM and CPM (FIG. 26)

Cleaning and Drying the System.

The procedure is the same as that for [¹⁸F]FNB

Preloading of Reagents and Solvents.

The procedure is substantially the same as that for [¹⁸F]FNB, except forthe reagents and cartridge described as follows. For the reagentconfiguration of the RDM, mannose triflate (25˜30 mg) in 1.5 ml ofanhydrous MeCN was loaded into reservoir #3, 2 ml of aq. HCl (1M) wasplaced in reservoir #5, 1 ml of H₂O was placed into reservoir #6 and 1ml of anhydrous MeCN was placed in reservoir #8. Reservoir #2, #4 and #7were empty. Reservoir #1 was reserved for loading ¹⁸F ion. For thereagent configuration of CPM, 10 ml of H₂O was placed in reservoir C;reservoir A and D were empty. The purification cartridge consisted of acation and anion resin mixed column, C18 and Al₂O₃ column preconditionedwith 10 ml of ethanol and H₂O in the corresponding position of CPM.

Production and Activation of ¹⁸F Ion.

The procedure is same as that for [¹⁸F]FNB.

Radiosynthesis and Purification of [¹⁸F]FDG.

Triflate precursor in MeCN in reservoir #3 of RDM was delivered into thereaction vial. The reaction mixture was heated at 100° C. for 4 min toperform the radiofluorination of the precursor to produce the labeledproduct [¹⁸F]F-TAG, the radio HPLC of this intermediate is shown in FIG.34. The heating was applied for additional 4 min to remove the solventMeCN. After cooling down to room temperature, 1.5 ml of aq. HCl solutionin reservoir #5 was delivered into the vial and heated at 100° C. for 10min to produce the hydrolyzed product (See FIG. 35 for radio-HPLC).After cooling down, The reaction mixture was transferred into reservoirC in CPM to dilute further. 2 ml of H₂O in reservoir #6 was deliveredinto the reaction vial to wash and obtain the residual reaction mixturewhich was then transferred into reservoir C in the CPM. The dilutedreaction mixture was passed through the purification cartridge and theelution was delivered to the product vial.

Quality control of [¹⁸F]FDG—The total activity of [¹⁸F]FDG solution wasmeasured by dose calibrator and its radiochemical purity was checked byradio-TLC for quality control (See FIG. 36). The sample was spotted onTLC plate, which was developed in mixed solvent of MeCN/H₂O (95/5) andscanned by radio-TLC scanner.

Example 9 Preparation of N-succinimidyl-4-[¹⁸F]fluoro benzoate([¹⁸F]SFB) in the Integrated System, Including RDM, MRM and CPM

The preparation of [¹⁸F]SFB follows the reaction scheme and thedescription given below:

System Configuration.

A 5 mL V-vial with PEEK adapter lid containing seven tubing ports wasinstalled in a microwave module, comprising a CEM microwave system andan auxiliary controller. Three outputs of an RDM (configured withdedicated outputs for each reagent) was connected to three of the portson the vial adapter. In this synthesis, not all reagents were pre-storedon the RDM; rather they were injected via tubing from outside theshielded environment when needed. Four ports of the vial adapter wereconnected to the microwave reactor auxiliary controller. This controllerperforms the following functions: open a vent when adding reagents,apply vacuum and nitrogen stream during evaporation, and apply pressureto transfer product out of reactor via the dip tube. The dip tube wasconnected to the input of the CPM for purification of the final product.Reservoir C (diluent) of the CPM was loaded with 8 mL of a 5 vol. %solution of acetic acid in water. Reservoir A (wash) was loaded with 10mL of the mixed solvent MeCN/H₂O (v/v 2:1). Reservoir D (eluent) wasfilled with 3 mL of diethyl ether. A Merck EN cartridge (200 mg,conditioned with 10 mL of ethanol and 10 mL of a 5 vol. % solution ofacetic acid in water) was installed.

Production and Activation of [¹⁸F]Fluoride.

An aliquot of aqueous [¹⁸F]fluoride solution (50˜100 μL, 1850˜3700 MBq)was added to Kryptofix 2.2.2 (10 mg, 26.6 μmol) and 13 μL of a 1 Mpotassium carbonate solution. The mixture was diluted with 0.9 ml of dryacetonitrile and transferred to the reactor via RDM position 1. Withvacuum applied, the solvent was evaporated at a microwave power of 20 Wfor 3 min. Azeotropic drying was repeated with addition of 1 mLacetonitrile through RDM position 1.

Preparation of 4-[¹⁸F]Fluorobenzoic Acid ([¹⁸F]3).

A solution of 1 (ca. 2.5 mg, 7.0 μmol) in dry DMSO (300 μL) was added tothe vial containing the dried [K⊂2.2.2][¹⁸F]F salt through RDM position2. With reaction vial sealed (all valves closed), stirring and aircooling activated, the reaction was completed in 1 min with a microwavepower of 50 W. A solution of potassium tert.-butoxide (ca. 10 mg, 89.1μmol) in DMSO (300 μL) was then added through RDM position 2. The second(deprotection) step of the reaction was carried out with the vialsealed, stirring and air cooling activated, under a microwave power of40 W for 1 min to yield [¹⁸F]3.

Preparation and Purification of N-Succinimidylester4-[¹⁸F]Fluorobenzoate ([¹⁸F]4).

To [¹⁸F]3 in DMSO a solution of TSTU (30 mg, 100.0 μmol) in acetonitrile(2.5 mL) was added through RDM position 3. Linkage of the succinimidylmoiety to [¹⁸F]3 was performed with air cooling and stirring at amicrowave power of 30 W for 2 min. The reaction mixture was transferredto the CPM for dilution. In the CPM, the product was caused to flowthrough the installed Merck EN cartridge, followed by the wash solution,and then nitrogen to dry the cartridge. The eluent was caused to flowthrough the cartridge to recover the product into a collection vial.After a synthesis time of 35˜40 min 370˜1110 MBq (RCY: 20˜30%) of n.c.a.N-succinimidyl-4-[¹⁸F]fluorobenzoate was produced (radiochemicalpurity>98%).

The various descriptions set forth above of the reaction module and theother modules used therewith, referred to as special function modules,and the components comprising the modules are provided as examplesthereof and are not intended to limit the various components of themodules, the arrangement of the modules in an assembled modular reactionsystem, the use of the modules or other supplemental components whichmade be added thereto. Further, while some of the examples show systemswith only one reaction module and one or more different special functionmodules it is contemplated that the modular chemical reaction systemincorporating features of the invention may include multiple reactionmodules as well as multiples of the various specialty modules. Forexample, the system can include one or more reaction modules, one ormore reagent storage and delivery modules, one or more purificationmodules, one or more quality control and analysis modules, one or morechemical transfer modules, one or more aliquotting modules, one or moreconcentrating or drying modules, one or more chemical concentratingmodules and one or more radiation counting modules. Further, it iscontemplated that additional special function modules may be added tothe system to further expand the capability of the system. Stillfurther, the various modules can be arranged to operate in series or inparallel as required to produce one or more exit streams of the desiredend product.

It is further contemplated that the reaction modules as well as thespecial function modules may include any desired sensors (i.e.,temperature, pressure, pH, radiation counters, etc) and analyticalprobes (i.e., IR, UV, specialty probes for various chemicalconstituents, etc) for conducting process analysis during a reaction inthe system as well as transmission means (i.e., hard wired, fiberoptics, telemetry, etc) for delivery of the sensor or probe output to aremotely located controller or monitor. Other components of the variousmodules can include, pumps, valves, stirring systems, liquid separationsystems or other components, all remotely controlled, as are typicallyrequired to conduct an automated chemical reaction. While each of thereaction modules and special function modules include the sensors,probes, and controllers necessary for independent operation of thatmodule, they are also configured so that all of the modules comprising areaction system, the components thereof and the functions thereof can bemonitored and controlled by a single system monitor and controller suchas provided by a general purpose computer.

It is also contemplated that one or more components of the reactionmodule are moveable, using a remote located control system to, forexample, allow the system operator to provide or remove heating orcooling to the reaction vessel or vial or to allow addition or removalof materials to the reaction vessel or vial. In a like manner movementof the components in the special function modules may be provide andcontrolled. Still further, movement of the various modules inrelationship to the other modules is also contemplated so that variousdifferent modules can be plugged together to provide the desiredassembled reaction system. Video capability is provided so thatoperation can be observed.

As set forth above, the assembly and operation of the modular chemicalproduct system and each the modules comprising that system arecontrolled, monitored, observed and recorded using various audio,visual, and electronic means so that the operation of the system canalso be reviewed and duplicated.

We claim:
 1. A method of performing radiochemistry synthesis comprising:providing a vial having an upper rim, the vial configured to hold one ormore reactants or reactant products of the radiochemistry synthesis;providing a reactor vessel having a heating element and athermally-conductive heat transfer block, the vial being containedwithin the thermally-conductive heat transfer block; providing astationary fluidic interface disposed above the reactor vessel and vialand comprising a plurality of elastomeric stoppers, wherein at least oneof the elastomeric stoppers comprises a completely sealed surface and atleast one of the elastomeric stoppers comprises a port fluidicallycoupled to tubing; providing a horizontal actuator coupled to thereactor vessel and configured to move the reactor vessel along asubstantially horizontal axis of motion; providing a vertical actuatorcoupled to the reactor vessel and configured to move the reactor vesselalong a substantially vertical axis of motion; providing a motioncontroller operatively coupled to the horizontal actuator and thevertical actuator; and wherein the motion controller operates inaccordance with a program to selectively seal the upper rim of the vialagainst the plurality of elastomeric stoppers.
 2. The method of claim 1,wherein the program is preprogrammed using a control unit associatedwith a computer.
 3. The method of claim 1, wherein the tubing isconnected to a purification unit and contents of the vial aretransferred from the vial to the purification unit.
 4. The method ofclaim 3, wherein a dip tube extends from the stationary fluidicinterface and into the vial when the upper rim of the vial is sealedagainst the plurality of elastomeric stoppers.
 5. The method of claim 1,wherein the tubing is connected to a source of vacuum and the source ofvacuum transfers contents out of the vial.
 6. The method of claim 1,wherein the tubing is connected to a source of inert gas, wherein thesource of inert gas is introduced into the vial.
 7. The method of claim1, wherein the tubing is connected to one or more reagent sources,wherein at least one reagent is introduced into the vial.
 8. The methodof claim 1, further comprising measuring the radioactivity of thereactant products of the radiochemistry synthesis with a radiationsensor.
 9. The method of claim 1, wherein the stationary fluidicinterface comprises a first elastomeric stopper in a first position thatincludes a conduit that extends through the first elastomeric stopperand second elastomeric stopper in a second position that comprises thecompletely sealed surface.
 10. The method of claim 9, wherein the secondelastomeric stopper comprises a chemically inert gasket.
 11. The methodof claim 1, wherein the stationary fluidic interface comprises a firstelastomeric stopper in a first position that includes tubing thatextends through the first elastomeric stopper, a second elastomericstopper in a second position that comprises the completely sealedsurface, and third elastomeric stopper that includes a dip tube thatextends through the third elastomeric stopper.
 12. The method of claim11, further comprising sealing the upper rim of the vial against thefirst elastomeric stopper and adding reagents into the vial.
 13. Themethod of claim 12, wherein the reagents comprise an aqueous F-18solution, a dry organic solvent, a phase transfer agent, and acarbonate.
 14. The method of claim 12, further comprising mixing thecontents of the vial by bubbling an inert gas in the vial.
 15. Themethod of claim 11, further comprising sealing the upper rim of the vialagainst the second elastomeric stopper and heating the contents of thevial.
 16. The method of claim 11, further comprising sealing the upperrim of the vial against the third elastomeric stopper and transferringthe contents of the vial out of the vial.